A rapid detection method for flavor additives in tea

Abstract

The invention relates to the technical field of fluorescent sensor construction, and provides a method for rapid detection of flavor additives in tea leaves. The method comprises: firstly, adding a carboxyl-modified mesoporous silica nanomaterial into a fluorescent molecule rhodamine solution, and shaking and mixing at room temperature to obtain a mesoporous silica nanomaterial-rhodamine complex; then adding a phosphate buffer solution containing a specific androgen receptor, and reacting in a shaker; after the reaction, centrifugal washing is performed, the supernatant is removed, the phosphate buffer solution is added to the precipitate, and ultrasonic vortex mixing is performed, and the washing is repeated three times; the washed precipitate is dispersed in the phosphate buffer solution to obtain a multifunctional mesoporous silica nanocomposite material; then, extracting oak moss, an illegal additive in the tea leaves, mixing the extract with the material for reaction; centrifugation and separation to obtain a supernatant; and finally detecting the supernatant with a fluorescence analyzer, and obtaining the oak moss concentration of the tea sample according to the linear relationship fitted by the standard sample.

Description

Technical Field

[0001] The invention relates to the technical field of fluorescent sensor construction, and more specifically to a method for quickly detecting flavor additives in tea. Background Art

[0002] Tea is one of the three major non-alcoholic beverages in the world today. Tea is loved by people all over the world for its pleasant taste, rich aroma and health benefits. However, there are many types of tea on the market with varying quality. Many unscrupulous merchants use inferior tea leaves and add flavor additives to cover up their authenticity. These additives are potentially harmful to the environment and human health.

[0003] Oakmoss is a kind of moss that grows on the trunk of an oak tree. It has a very unique fragrance, which is like turpentine and has the aroma of anise, bean, hay, etc. Due to its unique aroma and long-lasting fragrance, it can be used as a good fixative to enhance the fragrance of perfume and is widely used in the perfume industry. However, because oakmoss can cause skin pathological reactions such as allergic dermatitis in the human body, the International Fragrance Association (IFRA) restricts the use of oakmoss and stipulates that its addition to perfumes should not exceed 0.1%. However, some unscrupulous tea merchants are currently adding oakmoss to tea to increase the fragrance of tea. Oakmoss (Atraric acid), scientific name 2,4-dihydroxy 3,6-dimethylbenzoic acid methyl ester, has been found that large amounts of oakmoss tea or long-term drinking can cause irritability, vomiting, dizziness, liver and kidney damage, and convulsions. In addition, oakmoss is also a specific androgen receptor antagonist. Absorption by the human body may reduce the effect of androgens and cause metabolic disorders.

[0004] Aroma plays an important role in tea quality identification. For the evaluation of tea aroma quality, it is difficult for ordinary consumers to effectively identify and distinguish illegal aroma additives by observing the color, smelling the aroma, tasting the tea soup, etc., which has certain limitations. Nowadays, there are many methods for determining the aroma substances in tea by chemical methods, such as gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and high-performance liquid chromatography. These methods have high precision and high sensitivity, but the method cost is high, the steps are complicated, and they need to be carried out under the supervision of professionals.

[0005] Fluorescent biosensor is an efficient and accurate detection and analysis method, which mainly uses fluorescent dyes and fluorescent markers to detect the interaction and existence of biomolecules. It has been widely used in medicine, environmental protection, food testing, biomaterials and environmental monitoring. As a fast, efficient, simple and highly sensitive analytical technology, fluorescent biosensor has been widely used in medicine, environmental protection, food testing, biomaterials and environmental monitoring. It also shows great potential in the physical and chemical testing, quality evaluation and authenticity identification of tea and tea products. This technology provides a new idea for solving the problem of detecting illegal additives in tea.

[0006] In order to solve the above problems, a technical solution is now provided. Summary of the invention

[0007] In order to overcome the above-mentioned defects of the prior art, the present invention provides a method for rapid detection of flavor additives in tea. By combining a mesoporous silica nanomaterial carrier with a fluorescent biosensor, rapid quantitative detection of oak moss, an illegal additive in tea, is achieved, which solves the problems that the detection method of illegal additives in tea is single, the pretreatment and detection process are cumbersome, the detection site requirements are high, and the operator specialization level is high, which limits its application in oak moss detection.

[0008] To achieve the above object, the present invention provides the following technical solutions:

[0009] A rapid detection method for flavor additives in tea, using mesoporous silica as a carrier, androgen receptor as a specific recognition molecule, and rhodamine as a fluorescent signal molecule, comprising the following steps:

[0010] Step 1: adding the carboxyl-modified mesoporous silica nanomaterial to the fluorescent molecule rhodamine solution, and mixing by shaking at room temperature using an oscillator to obtain a mesoporous silica nanomaterial-rhodamine complex;

[0011] Step 2, adding a phosphate buffer solution containing a specific androgen receptor to the complex, and mixing and reacting on a shaker;

[0012] Step 3: After the mixing reaction is completed, centrifuge and wash, remove the supernatant, add phosphate buffer solution to the precipitate, perform ultrasonic vortex mixing, and repeat washing three times;

[0013] Step 4, dispersing the washed precipitate into a phosphate buffer solution to obtain a synthesized multifunctional mesoporous silica nanocomposite material;

[0014] Step 5, extracting the illegal additive oak moss in tea by using the QuEChERS method, mixing the tea extract with the multifunctional mesoporous silica nanomaterial, reacting in a shaker, and centrifuging to obtain a supernatant;

[0015] Step six, using a fluorescence analyzer to detect the fluorescence signal of the supernatant, and obtaining the oak moss concentration of the test tea sample based on the linear relationship fitted by the standard sample.

[0016] As a further solution of the present invention, in step 1, the mass of the carboxyl-modified mesoporous silica nanomaterial is 5 mg; the volume of the rhodamine solution is 500 μL, and the concentration is 5 mM; the rotation speed of the oscillator is 400 rpm, and the reaction time is 12 h.

[0017] As a further solution of the present invention, in step 2, the volume of the phosphate buffer solution containing androgen receptor is 500 μL, and the concentration is 10 μg / mL; the rotation speed of the shaker is 200 rpm, and the reaction time of the mixing reaction is 4 h.

[0018] As a further solution of the present invention, in step three, the centrifugal washing is carried out in a high-speed centrifuge at a centrifugal speed of 8000 rpm; and the volume of the phosphate buffer solution is 1 mL.

[0019] As a further solution of the present invention, in step 4, the volume of the phosphate buffer solution is 1 mL.

[0020] As a further solution of the present invention, in step five, the volume of the tea extract is 100 μL; the volume of the synthetic material is 100 μL; the shaking reaction conditions are temperature 37°C, speed 200 rpm, reaction time 2 h; the centrifugal condition is speed 8000 rpm.

[0021] As a further solution of the present invention, in step six, the fluorescence signal intensity of rhodamine in the detected supernatant is linearly related to the logarithm of the oak moss sample concentration; its linear response range is 0.1 ppb to 1 ppm, and the detection limit is 0.05 ppb.

[0022] As a further solution of the present invention, in step 5, the illegal additive oak moss in tea is extracted using the QuEChERS method, comprising the following steps:

[0023] Step S1, weighing tea leaves and grinding them into powder, adding an acetonitrile solution containing acetic acid to extract the additive oak moss from the tea leaves;

[0024] Step S2, separating the oak moss and the tea leaves by solid phase extraction;

[0025] Step S3, taking 2 mL of the upper layer liquid, concentrating it to near dryness using a nitrogen blower, and adding 1 mL of acetonitrile to obtain the tea extract.

[0026] As a further solution of the present invention, in step S1, the mass of the tea leaves is 2 g; the volume of the acetonitrile solution containing acetic acid is 15 mL, wherein the content of acetic acid is 1%; and the extraction process includes vigorous shaking for 15 minutes, followed by centrifugation at 4200 rpm for 5 minutes.

[0027] Compared with the prior art, the rapid detection method of flavor additives in tea of ​​the present invention has the following beneficial effects:

[0028] 1. The fluorescent biosensor of the present invention has high selectivity and specificity for oak moss detection, is less interfered by other substances in tea leaves, has low detection cost, and is quick and simple;

[0029] 2. The detection in the present invention does not require the reservation of a large-scale liquid phase mass spectrometer, and can be completed in a relatively short time using only a fluorescence analyzer, thus meeting the current growing demand for rapid detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The figure is a schematic flow chart of a rapid detection method of flavor additives in tea leaves according to the present invention;

[0031] Figure 2 Schematic diagram of fluorescent biosensor detection of oak moss, an additive in tea leaves;

[0032] Figure 3 This is a characterization diagram of mesoporous silica nanomaterials in a rapid detection method for flavor additives in tea of ​​the present invention;

[0033] Figure 4 This is a diagram of reaction condition optimization for a rapid detection method of flavor additives in tea leaves according to the present invention;

[0034] Figure 5 This is a performance diagram of a fluorescence quantitative sensor for a rapid detection method of flavor additives in tea leaves according to the present invention.

[0035] In the figure, Block: blocking; Atraric acid: oak moss; Release: release; Mesoporous silicananoparticles: mesoporous silica nanoparticles; Androgen receptor: androgen receptor; Rh6G: rhodamine; Relative frequency: relative frequency; Diameter: diameter; Zeta potential: Zeta potential; MSN: mesoporous silica nanoparticles; MSN-COOH: carboxyl-modified mesoporous silica nanoparticles; AR+MSN-COOH: androgen receptor-modified carboxyl mesoporous silica nanoparticles; FL Intensity (au): fluorescence intensity (arbitrary unit); Incubation time (h): incubation time (hour); Reaction time (h): reaction time (hour); Wavelength / nm: wavelength / nanometer; Blank: blank control; Log cocentration of AA (ng / mL): logarithm of oakmoss concentration (nanograms / milliliter); Concentration (ng / mL): concentration (nanograms / milliliter); AA: oakmoss; EGC: epigallocatechin-3-oxide; EGCG: epigallocatechin-3-oxide; PC: proanthocyanidins; SA: salicylic acid; CGA: chlorogenic acid; Teapolyphenols: tea polyphenols. DETAILED DESCRIPTION

[0036] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] Example 1

[0038] like Figure 1 The figure shows a schematic flow chart of a rapid detection method of flavor additives in tea leaves of the present invention, comprising the following steps:

[0039] Step 1: adding the carboxyl-modified mesoporous silica nanomaterial to the fluorescent molecule rhodamine solution, and mixing by shaking at room temperature using an oscillator to obtain a mesoporous silica nanomaterial-rhodamine complex;

[0040] Step 2, adding a phosphate buffer solution containing androgen receptor to the complex, and mixing and reacting on a shaker;

[0041] Step 3: After the mixing reaction is completed, centrifuge and wash, remove the supernatant, add phosphate buffer solution to the precipitate, perform ultrasonic vortex mixing, and repeat washing three times;

[0042] Step 4, dispersing the washed precipitate into a phosphate buffer solution to obtain a synthesized multifunctional mesoporous silica nanocomposite material;

[0043] Step 5, extracting the illegal additive oak moss in tea by using the QuEChERS method, mixing the tea extract with the multifunctional mesoporous silica nanomaterial, reacting in a shaker, and centrifuging to obtain a supernatant;

[0044] Step six, using a fluorescence analyzer to detect the fluorescence signal of the supernatant, and obtaining the oak moss concentration of the test tea sample based on the linear relationship fitted by the standard sample.

[0045] Example 2

[0046] A rapid detection method for flavor additives in tea, using androgen receptor as a specific recognition molecule and rhodamine as a fluorescent signal molecule, comprising the step of adding a carboxyl-modified mesoporous silica nanomaterial to a rhodamine solution, and mixing by shaking at room temperature using an oscillator to obtain a mesoporous silica nanomaterial-rhodamine complex.

[0047] In step 1 of the embodiment of the present invention, the mass of the carboxyl-modified mesoporous silica nanomaterial is 5 mg; the volume of the rhodamine solution is 500 μL, and the concentration is 5 mM; the rotation speed of the oscillator is 400 rpm, and the reaction time is 12 h.

[0048] The method for rapid detection of flavor additives in tea includes step 2, adding a phosphate buffer solution containing androgen receptor to the complex and mixing and reacting in a shaker.

[0049] In step 2 of the embodiment of the present invention, the volume of the phosphate buffer solution containing the androgen receptor is 500 μL; the rotation speed of the shaker is 200 rpm, and the reaction time of the mixing reaction is 4 h.

[0050] The method for rapid detection of flavor additives in tea includes step three, performing centrifugal washing after the mixing reaction is completed, removing the supernatant, adding phosphate buffer solution to the precipitate, performing ultrasonic vortex mixing, and repeating the washing three times.

[0051] In step three of the embodiment of the present invention, the centrifugal washing is performed in a high-speed centrifuge at a centrifugal speed of 8000 rpm; the volume of the phosphate buffer solution is 1 mL.

[0052] The method for rapid detection of flavor additives in tea includes step 4, dispersing the washed precipitate into a phosphate buffer solution to obtain a synthesized multifunctional mesoporous silica nanomaterial.

[0053] In step 4 of the embodiment of the present invention, the volume of the phosphate buffer solution is 1 mL.

[0054] The method for rapid detection of flavor additives in tea includes step five, using the QuEChERS method to extract the illegal additive oak moss in tea, mixing the tea extract with the multifunctional mesoporous silica nanomaterial, reacting in a shaker, and centrifuging to obtain a supernatant.

[0055] In step five of the embodiment of the present invention, the volume of the tea extract is 100 μL; the volume of the synthetic material is 100 μL; the shaking reaction conditions are temperature 37°C, speed 200 rpm, reaction time 2 h; the centrifugal condition is speed 8000 rpm.

[0056] In step 5 of the embodiment of the present invention, the QuEChERS method is used to extract the illegal additive oak moss in tea, which includes the following steps:

[0057] Step S1, weighing tea leaves and grinding them into powder, adding an acetonitrile solution containing acetic acid to extract the additive oak moss from the tea leaves;

[0058] Step S2, separating the oak moss and the tea leaves by solid phase extraction;

[0059] Step S3, taking 2 mL of the upper layer liquid, concentrating it to near dryness using a nitrogen blower, and adding 1 mL of acetonitrile to obtain the tea extract.

[0060] In step S1 of the embodiment of the present invention, the mass of the tea leaves is 2 g; the volume of the acetonitrile solution containing acetic acid is 15 mL, wherein the content of acetic acid is 1%; and the extraction process includes vigorous shaking for 15 minutes and then centrifugation at 4200 rpm for 5 minutes.

[0061] The method for rapid detection of flavor additives in tea includes step six, using a fluorescence analyzer to detect fluorescence signals of the supernatant, and obtaining the oak moss concentration of the test tea sample according to the linear relationship fitted by the standard sample.

[0062] In step six of the embodiment of the present invention, the fluorescence signal intensity of the rhodamine molecules in the detected supernatant and the logarithm of the oak moss sample concentration show a good linear relationship; the linear response range is 0.1 ppb to 1 ppm, and the detection limit is 0.05 ppb.

[0063] Example 3

[0064] In the embodiment of the present invention, a multifunctional mesoporous silica nanomaterial (MSN) is synthesized. First, 5 mg of carboxyl-modified mesoporous silica nanomaterial is added to 500 μL of 5 mM rhodamine (Rh6G), and the mixture is reacted on a shaker at 400 rpm for 12 h at room temperature, and the two are shaken and mixed; then 500 μL of phosphate buffer solution containing androgen receptor is added to MSN-Rh6G; then the mixture is mixed and reacted on a shaker at 200 rpm for 4 h; after the mixture is centrifuged and washed in a high-speed centrifuge at 8000 rpm, the supernatant is removed, 1 mL of phosphate buffer solution is added to the precipitate, and the mixture is ultrasonically vortexed and mixed, and the mixture is repeated three times; finally, the washed precipitate is dispersed in 1 mL of phosphate buffer solution (PBS) to obtain a multifunctional mesoporous silica nanocomposite material.

[0065] In the present invention, the synthesized multifunctional mesoporous silica nanomaterials are characterized, such as Figure 2 The figure shows a schematic diagram of fluorescent biosensor detection of oak moss, an additive in tea. First, the fluorescent signal molecule rhodamine is encapsulated in a carboxyl-modified mesoporous silica nanomaterial. Then, the androgen receptor is used as a specific recognition molecule to cover the surface of the mesoporous silica nanomaterial through electrostatic interaction, so that the fluorescent molecule cannot escape on its own. Because oak moss is a specific antagonist of the androgen receptor, the interaction between the two is stronger than the electrostatic force. After the addition of oak moss, AR combines with oak moss to release the fluorescent signal molecule Rh6G. By detecting the intensity of its fluorescent signal, the concentration of the added oak moss can be obtained according to the linear relationship.

[0066] Example 4

[0067] like Figure 3 Shown is a characterization diagram of mesoporous silica nanomaterials in a rapid detection method for flavor additives in tea of ​​the present invention.

[0068] In the embodiment of the present invention, the morphology of the mesoporous silica nanomaterial is characterized by transmission electron microscopy. The results show that the mesoporous silica nanomaterial is a round sphere with a regular shape and uniform dispersion, and the average particle size is about 110 nm.

[0069] Compared with the single mesoporous silica nanomaterial, the carboxyl-modified mesoporous silica nanomaterial in the embodiment of the present invention has a more negative Zeta potential, which proves that the carboxyl group is successfully modified on the surface of the mesoporous silica nanomaterial. After the androgen receptor is modified on the surface of the carboxyl-modified mesoporous silica nanomaterial by electrostatic interaction, the Zeta potential changes from negative to positive, indicating that the androgen receptor molecule is successfully modified on its surface.

[0070] Example 5

[0071] like Figure 4 Shown is a diagram of reaction condition optimization for a rapid detection method for flavor additives in tea leaves according to the present invention.

[0072] The reaction conditions mainly optimized in the examples of the present invention include the reaction time of androgen receptor (AR) encapsulating carboxyl-modified mesoporous silica nanomaterial (MSN-COOH) and the reaction time of oak moss sample and fluorescent sensor.

[0073] In the embodiment of the present invention, after a series of time gradient experiments, it was found that the fluorescence effect of the androgen receptor encapsulating the carboxyl-modified mesoporous silica nanomaterial was optimal when the time was about 4 h. If the time was too short, the androgen receptor could not completely encapsulate the carboxyl-modified mesoporous silica nanomaterial. If the time was too long, the stability of AR-MSN-COOH would decrease. For the reaction time of the tested tea samples, 2 h could meet the requirements.

[0074] Example 6

[0075] like Figure 5 Shown is a performance diagram of a fluorescence quantitative sensor for a rapid detection method of flavor additives in tea leaves according to the present invention.

[0076] In the embodiment of the present invention, by testing the concentration gradient of a series of standard samples, it is observed that there is a good linear relationship between the fluorescence signal intensity of the rhodamine molecule and the logarithm of the sample concentration, wherein the fitted linear equation is:

[0077] ;

[0078] In the formula, is the logarithm of the sample concentration, is the intensity of the fluorescence signal.

[0079] The correlation coefficient of the linear equation in the embodiment of the present invention is 0.993, the linear response range is 0.1 ppb-1ppm, and the detection limit is as low as 0.05 ppb.

[0080] In order to verify the specificity of the detection method in the examples of the present invention, various tea polyphenols in tea were used as negative controls for testing. The results showed that tea polyphenols did not significantly interfere with the detection of oak moss. Therefore, the sensor has excellent sensitivity and anti-interference performance, further proving that it can become a high-precision tool for oak moss detection.

[0081] Example 7

[0082] In the embodiment of the present invention, the QuEChERS method is used to extract the illegal additive oak moss in tea, including extraction, purification, concentration and analysis, and the specific steps are:

[0083] Step S1, accurately weigh 2 g of tea leaves, grind into powder, add 15 mL of acetonitrile solution containing 1% acetic acid, shake vigorously for 15 min, centrifuge at 4200 rpm for 5 min, and extract the additive oak moss from the tea leaves;

[0084] Step S2, separating the oak moss and the tea leaves by solid phase extraction;

[0085] Step S3, taking 2 mL of the upper layer liquid, concentrating it to near dryness using a nitrogen blower, adding 1 mL of acetonitrile to obtain the tea extract, and then testing it with fluorescence detection.

[0086] The fluorescence sensor developed in the embodiment of the present invention has similar results to those measured by liquid phase mass spectrometry, with high recovery rate and high accuracy. The results are shown in Table 1:

[0087]

[0088] Table 1. Comparison of the results measured by fluorescence sensor and liquid chromatography mass spectrometry

[0089] The above is only a specific implementation of the present application, but the protection scope of the present application is not limited thereto. Any person skilled in the art who is familiar with the present technical field can easily think of changes or substitutions within the technical scope disclosed in the present application, which should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be based on the protection scope of the claims.

[0090] Finally: The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A rapid detection method for flavor additives in tea, characterized in that: Mesoporous silica is used as a carrier, androgen receptor is used as a specific recognition molecule, and rhodamine is used as a fluorescent signal molecule, including the following steps: Step 1: adding the carboxyl-modified mesoporous silica nanomaterial to the fluorescent molecule rhodamine solution, and mixing by shaking at room temperature using an oscillator to obtain a mesoporous silica nanomaterial-rhodamine complex; Step 2, adding a phosphate buffer solution containing a specific androgen receptor to the complex, and mixing and reacting on a shaker; Step 3: After the mixing reaction is completed, centrifuge and wash, remove the supernatant, add phosphate buffer solution to the precipitate, perform ultrasonic vortex mixing, and repeat washing three times; Step 4, dispersing the washed precipitate into a phosphate buffer solution to obtain a synthesized multifunctional mesoporous silica nanocomposite material; Step 5, extracting the illegal additive oak moss in tea by using the QuEChERS method, mixing the tea extract with the multifunctional mesoporous silica nanocomposite material, reacting in a shaker, and centrifuging to obtain a supernatant; Step six, using a fluorescence analyzer to detect the fluorescence signal of the supernatant, and obtaining the oak moss concentration of the test tea sample based on the linear relationship fitted by the standard sample.

2. The rapid detection method of flavor additives in tea according to claim 1, characterized in that: In step 1, the mass of the carboxyl-modified mesoporous silica nanomaterial is 5 mg; the volume of the rhodamine solution is 500 μL, and the concentration is 5 mM; the rotation speed of the oscillator is 400 rpm, and the reaction time is 12 h.

3. The rapid detection method of flavor additives in tea according to claim 1, characterized in that: In step 2, the volume of the phosphate buffer solution containing the specific androgen receptor is 500 μL, and the concentration is 10 μg / mL; the rotation speed of the shaker is 200 rpm, and the reaction time of the mixing reaction is 4 h.

4. The rapid detection method of flavor additives in tea according to claim 1, characterized in that: In step 3, the centrifugal washing is performed in a high-speed centrifuge at a centrifugal speed of 8000 rpm; the volume of the phosphate buffer solution is 1 mL.

5. The rapid detection method of flavor additives in tea according to claim 1, characterized in that: In step 4, the volume of the phosphate buffer solution is 1 mL.

6. The rapid detection method of flavor additives in tea according to claim 1, characterized in that: In step 5, the volume of the tea extract is 100 μL; the shaking reaction conditions are temperature 37° C., rotation speed 200 rpm, and reaction time 2 h; and the centrifugal condition is rotation speed 8000 rpm.

7. The rapid detection method of flavor additives in tea according to claim 1, characterized in that: In step 6, the fluorescence signal intensity of rhodamine in the detected supernatant showed a good linear relationship with the logarithm of the oak moss sample concentration; Its linear response range is from 0.1 ppb to 1 ppm, and its detection limit is 0.05 ppb.

8. The rapid detection method of flavor additives in tea according to claim 1, characterized in that: In step 5, the QuEChERS method is used to extract the illegal additive oak moss in tea, which includes the following steps: Step S1, weighing tea leaves and grinding them into powder, adding an acetonitrile solution containing acetic acid to extract the additive oak moss from the tea leaves; Step S2, separating the oak moss and the tea leaves by solid phase extraction; Step S3, taking 2 mL of the upper layer liquid, concentrating it to near dryness using a nitrogen blower, and adding 1 mL of acetonitrile to obtain the tea extract.

9. A rapid detection method for flavor additives in tea according to claim 8, characterized in that: In step S1, the mass of the tea leaves is 2 g; the volume of the acetonitrile solution containing acetic acid is 15 mL, wherein the content of acetic acid is 1%; the extraction process includes vigorous shaking for 15 minutes and then centrifugation at 4200 rpm for 5 minutes.

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